In situ reprogramming to transdifferentiate fibroblasts into cardiomyocytes using adenoviral vectors: Implications for clinical myocardial regeneration

Megumi Mathison, Vivek P. Singh, Maria J. Chiuchiolo, Deepthi Sanagasetti, Yun Mao, Vivekkumar B. Patel, Jianchang Yang, Stephen M. Kaminsky, Ronald Crystal, Todd K. Rosengart

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Objective The reprogramming of cardiac fibroblasts into induced cardiomyocyte-like cells improves ventricular function in myocardial infarction models. Only integrating persistent expression vectors have thus far been used to induce reprogramming, potentially limiting its clinical applicability. We therefore tested the reprogramming potential of nonintegrating, acute expression adenoviral (Ad) vectors. Methods Ad or lentivirus vectors encoding Gata4 (G), Mef2c (M), and Tbx5 (T) were validated in vitro. Sprague-Dawley rats then underwent coronary ligation and Ad-mediated administration of vascular endothelial growth factor to generate infarct prevascularization. Three weeks later, animals received Ad or lentivirus encoding G, M, or T (AdGMT or LentiGMT) or an equivalent dose of a null vector (n = 11, 10, and 10, respectively). Outcomes were analyzed by echocardiography, magnetic resonance imaging, and histology. Results Ad and lentivirus vectors provided equivalent G, M, and T expression in vitro. AdGMT and LentiGMT both likewise induced expression of the cardiomyocyte marker cardiac troponin T in approximately 6% of cardiac fibroblasts versus <1% cardiac troponin T expression in AdNull (adenoviral vector that does not encode a transgene)-treated cells. Infarcted myocardium that had been treated with AdGMT likewise demonstrated greater density of cells expressing the cardiomyocyte marker beta myosin heavy chain 7 compared with AdNull-treated animals. Echocardiography demonstrated that AdGMT and LentiGMT both increased ejection fraction compared with AdNull (AdGMT: 21% ± 3%, LentiGMT: 14% ± 5%, AdNull: −0.4% ± 2%; P < .05). Conclusions Ad vectors are at least as effective as lentiviral vectors in inducing cardiac fibroblast transdifferentiation into induced cardiomyocyte-like cells and improving cardiac function in postinfarct rat hearts. Short-term expression Ad vectors may represent an important means to induce cardiac cellular reprogramming in humans.

Original languageEnglish
Pages (from-to)329-339.e3
JournalJournal of Thoracic and Cardiovascular Surgery
Volume153
Issue number2
DOIs
Publication statusPublished - 1 Feb 2017
Externally publishedYes

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Cardiac Myocytes
Lentivirus
Regeneration
Fibroblasts
Troponin T
Echocardiography
Ventricular Myosins
Ventricular Function
Myosin Heavy Chains
Transgenes
Vascular Endothelial Growth Factor A
Ligation
Sprague Dawley Rats
Myocardium
Histology
Cell Count
Myocardial Infarction
Magnetic Resonance Imaging

Keywords

  • adenoviral vector
  • gene therapy
  • in situ cardiac reprogramming

ASJC Scopus subject areas

  • Surgery
  • Pulmonary and Respiratory Medicine
  • Cardiology and Cardiovascular Medicine

Cite this

In situ reprogramming to transdifferentiate fibroblasts into cardiomyocytes using adenoviral vectors : Implications for clinical myocardial regeneration. / Mathison, Megumi; Singh, Vivek P.; Chiuchiolo, Maria J.; Sanagasetti, Deepthi; Mao, Yun; Patel, Vivekkumar B.; Yang, Jianchang; Kaminsky, Stephen M.; Crystal, Ronald; Rosengart, Todd K.

In: Journal of Thoracic and Cardiovascular Surgery, Vol. 153, No. 2, 01.02.2017, p. 329-339.e3.

Research output: Contribution to journalArticle

Mathison, Megumi ; Singh, Vivek P. ; Chiuchiolo, Maria J. ; Sanagasetti, Deepthi ; Mao, Yun ; Patel, Vivekkumar B. ; Yang, Jianchang ; Kaminsky, Stephen M. ; Crystal, Ronald ; Rosengart, Todd K. / In situ reprogramming to transdifferentiate fibroblasts into cardiomyocytes using adenoviral vectors : Implications for clinical myocardial regeneration. In: Journal of Thoracic and Cardiovascular Surgery. 2017 ; Vol. 153, No. 2. pp. 329-339.e3.
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abstract = "Objective The reprogramming of cardiac fibroblasts into induced cardiomyocyte-like cells improves ventricular function in myocardial infarction models. Only integrating persistent expression vectors have thus far been used to induce reprogramming, potentially limiting its clinical applicability. We therefore tested the reprogramming potential of nonintegrating, acute expression adenoviral (Ad) vectors. Methods Ad or lentivirus vectors encoding Gata4 (G), Mef2c (M), and Tbx5 (T) were validated in vitro. Sprague-Dawley rats then underwent coronary ligation and Ad-mediated administration of vascular endothelial growth factor to generate infarct prevascularization. Three weeks later, animals received Ad or lentivirus encoding G, M, or T (AdGMT or LentiGMT) or an equivalent dose of a null vector (n = 11, 10, and 10, respectively). Outcomes were analyzed by echocardiography, magnetic resonance imaging, and histology. Results Ad and lentivirus vectors provided equivalent G, M, and T expression in vitro. AdGMT and LentiGMT both likewise induced expression of the cardiomyocyte marker cardiac troponin T in approximately 6{\%} of cardiac fibroblasts versus <1{\%} cardiac troponin T expression in AdNull (adenoviral vector that does not encode a transgene)-treated cells. Infarcted myocardium that had been treated with AdGMT likewise demonstrated greater density of cells expressing the cardiomyocyte marker beta myosin heavy chain 7 compared with AdNull-treated animals. Echocardiography demonstrated that AdGMT and LentiGMT both increased ejection fraction compared with AdNull (AdGMT: 21{\%} ± 3{\%}, LentiGMT: 14{\%} ± 5{\%}, AdNull: −0.4{\%} ± 2{\%}; P < .05). Conclusions Ad vectors are at least as effective as lentiviral vectors in inducing cardiac fibroblast transdifferentiation into induced cardiomyocyte-like cells and improving cardiac function in postinfarct rat hearts. Short-term expression Ad vectors may represent an important means to induce cardiac cellular reprogramming in humans.",
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T1 - In situ reprogramming to transdifferentiate fibroblasts into cardiomyocytes using adenoviral vectors

T2 - Implications for clinical myocardial regeneration

AU - Mathison, Megumi

AU - Singh, Vivek P.

AU - Chiuchiolo, Maria J.

AU - Sanagasetti, Deepthi

AU - Mao, Yun

AU - Patel, Vivekkumar B.

AU - Yang, Jianchang

AU - Kaminsky, Stephen M.

AU - Crystal, Ronald

AU - Rosengart, Todd K.

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N2 - Objective The reprogramming of cardiac fibroblasts into induced cardiomyocyte-like cells improves ventricular function in myocardial infarction models. Only integrating persistent expression vectors have thus far been used to induce reprogramming, potentially limiting its clinical applicability. We therefore tested the reprogramming potential of nonintegrating, acute expression adenoviral (Ad) vectors. Methods Ad or lentivirus vectors encoding Gata4 (G), Mef2c (M), and Tbx5 (T) were validated in vitro. Sprague-Dawley rats then underwent coronary ligation and Ad-mediated administration of vascular endothelial growth factor to generate infarct prevascularization. Three weeks later, animals received Ad or lentivirus encoding G, M, or T (AdGMT or LentiGMT) or an equivalent dose of a null vector (n = 11, 10, and 10, respectively). Outcomes were analyzed by echocardiography, magnetic resonance imaging, and histology. Results Ad and lentivirus vectors provided equivalent G, M, and T expression in vitro. AdGMT and LentiGMT both likewise induced expression of the cardiomyocyte marker cardiac troponin T in approximately 6% of cardiac fibroblasts versus <1% cardiac troponin T expression in AdNull (adenoviral vector that does not encode a transgene)-treated cells. Infarcted myocardium that had been treated with AdGMT likewise demonstrated greater density of cells expressing the cardiomyocyte marker beta myosin heavy chain 7 compared with AdNull-treated animals. Echocardiography demonstrated that AdGMT and LentiGMT both increased ejection fraction compared with AdNull (AdGMT: 21% ± 3%, LentiGMT: 14% ± 5%, AdNull: −0.4% ± 2%; P < .05). Conclusions Ad vectors are at least as effective as lentiviral vectors in inducing cardiac fibroblast transdifferentiation into induced cardiomyocyte-like cells and improving cardiac function in postinfarct rat hearts. Short-term expression Ad vectors may represent an important means to induce cardiac cellular reprogramming in humans.

AB - Objective The reprogramming of cardiac fibroblasts into induced cardiomyocyte-like cells improves ventricular function in myocardial infarction models. Only integrating persistent expression vectors have thus far been used to induce reprogramming, potentially limiting its clinical applicability. We therefore tested the reprogramming potential of nonintegrating, acute expression adenoviral (Ad) vectors. Methods Ad or lentivirus vectors encoding Gata4 (G), Mef2c (M), and Tbx5 (T) were validated in vitro. Sprague-Dawley rats then underwent coronary ligation and Ad-mediated administration of vascular endothelial growth factor to generate infarct prevascularization. Three weeks later, animals received Ad or lentivirus encoding G, M, or T (AdGMT or LentiGMT) or an equivalent dose of a null vector (n = 11, 10, and 10, respectively). Outcomes were analyzed by echocardiography, magnetic resonance imaging, and histology. Results Ad and lentivirus vectors provided equivalent G, M, and T expression in vitro. AdGMT and LentiGMT both likewise induced expression of the cardiomyocyte marker cardiac troponin T in approximately 6% of cardiac fibroblasts versus <1% cardiac troponin T expression in AdNull (adenoviral vector that does not encode a transgene)-treated cells. Infarcted myocardium that had been treated with AdGMT likewise demonstrated greater density of cells expressing the cardiomyocyte marker beta myosin heavy chain 7 compared with AdNull-treated animals. Echocardiography demonstrated that AdGMT and LentiGMT both increased ejection fraction compared with AdNull (AdGMT: 21% ± 3%, LentiGMT: 14% ± 5%, AdNull: −0.4% ± 2%; P < .05). Conclusions Ad vectors are at least as effective as lentiviral vectors in inducing cardiac fibroblast transdifferentiation into induced cardiomyocyte-like cells and improving cardiac function in postinfarct rat hearts. Short-term expression Ad vectors may represent an important means to induce cardiac cellular reprogramming in humans.

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KW - gene therapy

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